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Biofuel from D-Xylose - the Second Most Abundant Sugar

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Biofuel from D-Xylose - the Second Most Abundant Sugar GENERAL I ARTICLE Biofuel from D-xylose - the Second Most Abundant Sugar Anil Lachke In the biosphere we find cellulose and hemicellulose as the major polysaccharides. On acid or enzymatic hydrolysis, D-glucose is produced from cellulose and D-xylose is pro­ duced from xylans as the major sugar in the hydrolysate. Initially it was believed that yeasts do not fermentD-xylose to ethanol although many are capable of producing xylitol. Anil Lachke is a scientist Twenty years ago, a few yeasts that could convertD-xylose in the Division of to ethanol were found. Ethanol is viewed as a potential fuel Biochemical Sciences of that is available from biomass and hence new methods to National Chemical generate ethanol from hitherto inaccessible sources are Laboratory, Pune. His major research interests gaining importance. Biotechnology for efficient utilization include pentose metabo- of lignocellulose wastes as fuels relies on the utilization of lism in yeasts, biotechnol­ both the cellulosic as well as hemicellulosic portions of the ogy for biomass utiliza­ biomass. In this article, conversion of xylose into ethanol tion and biodeinking for recycling of waste paper. is discussed. He takes special interest in popularization of Henry Ford was once asked about the fate of his automobile science and technology. business if the petroleum supplies of the world run out. He He likes Indian classical exclaimed, "we can get fuel from the sumac by road side or from music, yoga and drawing. weeds, sawdust almost anything. There is fuel in every bit ofvegetable matter that can be fermented .... A nd it remains for someone to find out how this fuel can be produced commercially ... Better fuel at a cheaper price than we now know." The optimism shown by Henry Ford must be appreciated. Many countries are showing increasing interest in obtaining a large fraction of their energy needs from biomass. More than 25% of the cars in Brazil run on gasohol (Box 1) - a mixture of petrol and alcohol. They produce alcohol from molasses. Cur­ rent scientific opinion seems to favour the view that the world's Keywords fossil fu~l resources are steadily diminishing. Alternative sources D-xylose, biofuel. of energy must therefore be investigated urgently. There is no ________,AAflAA" ______ __ 50 v V VV V v RESONANCE , May 2002 GENERAL I ARTICLE Box t. Gasohol- An Alternative Fuel AlcOhol was identified as a possible replacement fuel during 1920's when there was a growing demand for gasoline and potential shortage of oil. Many automobile giants like General Motors and Ford, predicted that alcohol could be a viable fuel for vehicles. A blend of gasoline and alcohol that is referred to as 'gasohol' was tried out in various cars. However, gasohol got a bad name because of excess use or wrong choice of alcohol. Ofthe two types of alcohols that could be used, ethyl alcohol and methyl alcohol can be used by itselfas a fuel but when mixed with gasoline, it is limited to 10% by volume. Methyl alcohol is very corrosive and this limits its use in the mixture. The heat content of a litre of alcohol is less than gasoline and hence more alcohol is required to achieve the same power levels as that of gasoline. The penalty is decreased fuel economy and lower driving range without installing bigger fuel tanks. Another disadvantage of alcohol as a fuel is its higher volatility. During hot weather driving, the fuel vaporises easily and vapour lock in the engine can occur. This can make the engine run rough or even prevent it from running. An advantage of using alcohol as a fuel is that it mixes easily with water and prevents ice formation in cold weather. It also has a high octane rating than gasoline, which allows engine compression ratios to be increased and ignition timing to be advanced for better performance. The successes in development of other alternate energy sources may limit the use of alcohol as a fuel, but there has been some work in using alcohol in conjunction with fuel cells to produce power for the future vehicles. Alcohol has been used as a fuel for centuries and it may well be the fuel of the future too. Currently, there is only one major fuel company in Canada, Mohawk, supplying fuel with alcohol in it. Brazil is also supplier and user of gasohol for a number of years. doubt that in the long run we have to tum to renewable sources of food and energy. Over 150 billion tonnes of organic substances are photosynthe­ sized annually which consists of three major constituents, namely cellulose, hemicellulose (Box 2) and lignin with an average proportion of 4:3:3. The xylan (hemicellulose) content of the hardwood and the softwood lies in the range of 20-25% and 7-12%, respectively. The incentives for energy recovery from wastes are most attrac­ ti ve in the rural areas of India. Lignocellulose as a raw material Over 150 billion for bioconversion has a role in energy production; it can prevent tonnes of organic deterioration of the environment, and facilitate waste manage­ substances are ment (Figure 2). Large quantities of lignocellulosic materials in photosynthesized the form of agricultural and forest residues are available in annually. -R-ES-O-N--A-N-C-E--I -M-O-Y--2-0-0-2--------------~~------------------------------~- GENERAL I ARTICLE Box 2. Hemicellulose Plants contain about 6 x 1011 tonnes of hemicellulose and about 3 x 10 10 tonnes are photosynthesized annually by higher plants to make over one third of their dry weight. The close association of hemicelluloses with cellulose and lignin confer rigidity to the plant cell wall. The hemicelluloses are composed of a linear as well as branched hetero- and homopolymers of pentosans (D-xylose and L-arabinose) and hexosans (D-glucose, D-mannose and D-galactose). Unlike cellulose hemicelluloses show structural variability (Figure I). As compared to cellulose, hemicelluloses are low molecular weight polymers with a degree of polymer­ ization of around 200. Native hemicelluloses like xylan is composed of 85-90% D-xylose and a small amount of L-arabinose and traces of glucuronic acid. HemiceUuloses have more branches and are less crystalline than cellulose. The glycosidic linkages between the anhydro-D-xylose residues in xylan are more susceptible to acid or enzymatic hydrolysis than linkages between anhydro-D-glucose residues in cellulose. As a result, pentose sugars can be obtained readily in hydrolysates with sufficient yields from hemicelluloses. Considering the complex structure of hemicelluloses, a diverse set of enzymes is necessary for its hydrolysis to form soluble pentose sugars. These include: Endo-l, 4-fJ-D-xylanase, Exo- 1,4-p-xylanase, 1,4-fJ-D-xylosidase and a-L-arabinofuranosidase. Figure 1. @ G) OH OH(j) H~2C HO~OH HO 0 O~rf HO o 0 HO HOH2C HO @ ~HOH2~ D-GLUCOPYRANOSE CELLOBIOSE UNIT RING CELLULOSE P(1~ 4)-D-XYLOPYRANOSE LINKAGE OH a-(1~3)- L-ARABINOFURANOSE LINKAGE '" __ O~O~O o 000 O~-\° -0-- HO~~ AcO~ OH H;?O HO OH 0 OH OAc D-XYLOPYRANOSE RING 0 HOOC OH a-(1----. 2) - 4-0-METHYL-D-GLUCURONIC Ac- Acetyl group ACID LINKAGE MeO HO XYLAN ________LAAAAAA __------ 52 v V VV V v RESONANCE I May 2002 GENERAL I ARTICLE Figure 2. Xylose Glucose Mannose, Arabinose Galactose • Energy (ATP) • Fuel • Energy (ATP) • Chemicals • Protein (SCP) • Chemicals • Proteins(SCP) • Solvents • Fermentation • Solvents • Chemicals Products • Beverages • Xylitol • Chemicals • Energy (ATP) • Fructose • Petrochemical Synthesis • Food! Feed India. For example, India's production of wheat straw and rice straw is over 200 mlnion tonnes per year. Although straw has low protein content it is rich in fermentable carbohydrates Also, hemicellulose rich waste materials like corncob or coconut pith or coconut shell are valuable renewable resources (Tables I and 2). In addition, cultivation of cereals generates a lot of waste residues which is underutilized. Hydrolysis of Cellulosics The ultimate product of cellulose hydrolysis is glucose; a well­ known fermentable six-carbon sugar while the hemicellulosic fraction gives D-xyJpse, a five-carbon sugar. Thus, D-xylose is • the second most abundant sugar in nature and a potential India's production feedstock for generating food and fuel. Cost analysis has indi-· of wheat straw and cated that economically viable bioconversion processes require rice straw is over quantitative conversion of cellulosic as well as herni-cellulosic 200 million tonnes sugars. Hemicellulosic sugars (D-xylose) can be obtained more per year. -R-ES-O---NA-N-C-E---IM-a-Y--2-00-2---------------~-------------------------------5-3 GENERAL I ARTICLE Table 1. Composition of BIOMASS CELLULOSE HEMI- LIGNIN selected Iigno-celluloslc CELLULOSE materials. (% Dry weight - subject to Rice straw 37 24 14 variation.) Wheat straw 31 29 18 Bamboo 40 20 20 Subabul 33 20 NA Mesta wood 35 18 NA readily with better yield of 80-90% from xylan by acid or enzy­ matic hy~rolysis than D-glucose from cellulose. Utilization of D-xylose for the commercial production of valuable chemicals like ethanol, acetic acid, 2, 3-butanediol, acetone, isopropanol and n-butanol using microorganisms is thus important for en­ hancing the economic viability of lignocellulose utilization. For a long time, it was believed that yeast could not ferment D­ xylose although many strains can ferment D-xylulose, the keto isomer ofD-xylose. In 1914, a famous microbiologist Kluyver in his PhD thesis mentioned that, " ... only a small amount ofxylose was available. Therefore, I did not routinely include xylose in my fermentation experiments. Although many fungi can assimilate this sugar there are so many reports on their inability to ferment that omission ofxylose was not considered to be serious.
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